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  #1  
Old 12-30-2018, 11:33 AM
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Default Vegetative propagation of cannabis by stem cuttings... a study

some interesting information...

Vegetative propagation of cannabis by stem cuttings:
effects of leaf number, cutting position, rooting hormone
and leaf tip removal


direct link:
https://tspace.library.utoronto.ca/b...-2018-0038.pdf
or
https://tspace.library.utoronto.ca/handle/1807/90574

abstract:

This study evaluated the influence of the several factors and their interactive effects on propagation success using stem cuttings of cannabis (Cannabis sativa L.). Factors included: (i) Leaf number (two or three) (ii) leaf tip removal (1/3 of leaf tips removed) (iii) basal/apical position of stem cutting on the stock plant, and (iv) rooting hormone (0.2% indole-3-butyric (IBA) acid gel or 0.2% willow (Salix alba L.) extract gel). Cuttings were placed in a growth chamber for twelve days then assessed on their rooting success rate and root quality using a relative root quality scale. The IBA gel delivered a 2.1 times higher rooting success rate and 1.6 times higher root quality than the willow extract. Removing leaf tips reduced rooting success rate from 71% to 53% without influencing root quality. Cuttings with three leaves had 15% higher root quality compared to those with two, but leaf number did not influence rooting success rate. Position of cutting had little effect on rooting success or quality. To achieve maximum rooting success and root quality, cuttings from either apical or basal positions should have at least three fully expanded, uncut leaves and the tested IBA rooting hormone is preferential to the willow-based product.
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Last edited by Efesto; 12-30-2018 at 11:52 PM.
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  #2  
Old 12-31-2018, 06:47 PM
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There's some solid proof finally.
I've been telling idiots for years not to mess with clones anymore than absolutely required.
Those self same idiots still clip the leaf tips from clones.....and still say that it works better.

The same idjits claim that cropping the lower leaves on a plant increases bud size.
The frickin idiots still don't understand that it lowers the amount of photosynthesis area of the plant and decreases the ability of the bud growth they are looking for by using the stupid method in the first place.

Ah well, superstitions are not my thing.
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Old 12-31-2018, 11:24 PM
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Amusing. I never cut my leaf tips and I use Rootone on my cuttings. I typically get plantable rooted clones within 10 days in my home made $1 Store cloner with an aquarium pump and air stone (so called bubble cloner). I tried willow bark water last year but it just added bacteria and I did not get any better results than when using Rootone alone in plain water. I also use a heat mat and keep the water in the cloner at 77 deg F. I use a T-8 5100k bulbs for a light source and I keep the rooting area dark when rooting in water.

I would be interested in seeing what the best pH is for rooting. The hydro guys say a really acidic soup is best, but I use 6.8 pH well water.
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Old 01-03-2019, 11:41 PM
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Quote:
Originally Posted by n2ishun View Post
The same idjits claim that cropping the lower leaves on a plant increases bud size...

I dare not even imagine what you think of growers who use the "schwazzing method"... ahaha lol

....

I copy and paste the study to fix it in the forum
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Last edited by Efesto; 01-04-2019 at 01:01 AM.
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Old 01-04-2019, 12:55 AM
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INTRODUCTION

Cannabis (Cannabis sativa L.) production for legal markets in North America, including both medical and recreational, is quickly becoming a profitable industry. North American spending on legal cannabis was estimated at 6.7 billion USD in 2016 and is projected to reach 21.6 billion by 2021 (ArcView Market Research 2017).
Cannabis is an annual herbaceous species which has been widely cultivated and used as a medicinal plant since ~ 2800 BCE (Russo 2007). Its medicinal value is attributed mainly to a group of secondary metabolites called cannabinoids which are concentrated mostly in the essential oils of unfertilized female cannabis flowers (Potter 2014). Cannabis cultivation and possession were outlawed in the United States in 1971 and much of the world followed suit soon after (Potter 2009). Since then, some countries including Canada and The Netherlands have relaxed their regulations and implemented programs allowing access to cannabis for medicinal purposes. In these programs, strict safety standards are enforced to control the quality of cannabis being distributed to patients; however, there is little guidance for growers regarding horticultural management. For growers, horticultural guides and online resources are available but few are based on peer-reviewed scientific research (Potter 2009; Caplan et al. 2017a; Caplan et al. 2017b)

Based on our communications with Canadian medicinal cannabis producers and recent reviews on the state of global cannabis production (Leggett 2006; Potter 2014; Farag and Kayser 2015) modern day cannabis production occurs primarily in controlled environments using artificial lighting and either soilless growing substrates (Caplan et al. 2017a; Caplan et al. 2017b) or solution culture. Further, some medical cannabis growers favor organic production practices since consumers and regulating bodies often demand pesticide-free cannabis. Cannabis is propagated by seed (Potter 2009; Farag and Kayser 2015), vegetative stem cuttings (Coffman and Gentner 1979; Potter 2009) and in-vitro propagation (Lata et al. 2009a, 2009b, 2011).
Propagation using vegetative stem cuttings is often preferred by cannabis growers. It is a low-cost method that delivers genetically uniform plants with consistent rates of growth and cannabinoid production when compared to propagation from seed (Coffman and Gentner 1979; Potter 2009).

To our knowledge, no peer-reviewed research exists on optimizing propagation by stem cuttings in cannabis; however, this method has been investigated in other economically important species such as Pisum sativum L. (Eliasson, 1978), Lippia javanica L. (Soundy et al. 2008), some timber crops (LeBude et al., 2004; Ofori et al., 1996) and some ornamental nursery crops (Grange and Loach 1985). The primary goal of propagation by stem cuttings is to facilitate the formation of adventitious roots. Several factors have been identified which support adventitious rooting in vegetative stem cuttings (Hartmann et al. 2002). Some of these include: leaf area (or leaf number), cutting position on the stock plant, the use of rooting hormones, lighting, rooting medium, water status and mineral nutrition. The present study focused on the first three of these factors.

Leaves act as sources of photosynthate for cuttings which is important for successful rooting.
Increased leaf area and/or number may improve rooting success rate and formation of adventitious roots in cuttings (Leakey and Coutts, 1989; Ofori et al., 1996). Leaves also stimulate rooting as sources of rooting co-factors and endogenous auxin (Haissig 1974). Conversely, greater leaf area and/or number provide a larger surface area for evapotranspiration and evapotranspirative water loss which may negatively affect rooting success rate (Davis and Potter 1989). A reduction in leaf area may reduce evapotranspiration-induced stress (Leakey and Coutts 1989; Ofori et al. 1996) or avoid crowding in the propagation environment.

(Aminah et al. 1997). A common practice in modern day cannabis productions, based on our communications with Canadian medicinal cannabis producers and gray resources (Cervantes 2006), is to keep 2 to 3 leaves on each cutting and to remove about 1/3 of the leaf tips. The optimal leaf number on stem cuttings varies between species (Machida et al. 1977; Aminah et al. 1997; Alves et al. 2016) so species-specific evaluations are necessary.

The ability of stem cuttings to form adventitious roots often depends on the maturity of the stock plant. Cuttings from juvenile plants generally have improved rooting over those from mature plants (Altamura 1996). Juvenile plant material sometimes has a higher content of endogenous auxins and other rooting promoters compared to mature material (Husen and Pal 2006). This difference is evident in a number hardwood species such as oak (Morgan and McWilliams 1976), teak (Husen and Pal 2006) and American elm (Schreiber and Kawase 1975). Further, in hardwood cuttings, maturity often varies by cutting position on the stock plant; stems from more basal regions often retain juvenile characteristics and have improved capacity to form
adventitious roots (Hackett 1970). There is limited information and mixed findings on the effects
of cutting position on adventitious rooting in softwood and herbaceous plants. In Schefflera arboricola, softwood cuttings from more basal regions had lower rooting success rate and number of roots (Hansen 1986) while in fever tea (Lippia javanica L.), cutting position had no effect on rooting success (Soundy et al. 2008).

It is well documented that treating the basal portions of stem cuttings in synthetic auxins such as indole-3-butyric acid (IBA) can improve rooting success rate, increase the speed of rooting and increase the quantity of adventitious roots (Hartmann et al. 2002). In organic production, synthetic auxins such as IBA are often not permitted; thus, alternatives hormones or techniques
are used to improve rooting success rate and quality. Willow (Salix alba L.) shoot extract is an naturally-derived alternative to synthetic auxins and has been used successfully as natural rooting hormone for mung bean (Vigna radiata L.) cuttings (Arena et al. 1997) but without effect on olive (Olea europaea L.) cuttings (Al-Amad and Qrunfleh 2016) or willow (Kawase 1964) cuttings. Currently there is no peer-reviewed literature on any of the factors described above on cannabis.

The objective of the present study was to evaluate the influence of the following factors and their interactive effects on propagation using stem cuttings in cannabis: (i) number of leaves, (ii) leaf tip removal, (iii) basal/apical position of stem cutting, and (iv) type of rooting hormone.

MATERIALS AND METHODS

Stock Plant Conditions

Stock (mother) plants were maintained under 18-hour photoperiod with a mean canopy-level light intensity at 105 molm-s- (s.d. 61.2 molm-s-) using ceramic metal halide 3100K lamps (Philips Lighting, Markham, ON, Canada). Temperature (day/night) was maintained at 20˚C (s.d. 0.03˚C), air relative humidity (RH; day/night) was maintained at 63% (s.d. 2.3) and carbon dioxide (CO2) concentration (day/night) was maintained at 646 ppm (s.d. 59.7).
Stock plants were potted in 12.5 L air pruning pots (306 mm diameter 275 mm height; Caledonian Tree Company Ltd., Pathhead, United Kingdom) containing a custom blended organic growing substrate (60% sphagnum peatmoss and 40% bulk coconut coir; Premier Tech, Rivire-du-Loup, QC, Canada). The stock plants were 10 months-old and had between 20 and 25
nodes on their main stems. The plants were fertigated as needed using Nutri Plus Organic Grow liquid organic fertilizer (4.0N1.3P1.7K; Nutri Plus; EZ-GRO Inc., Kingston, ON, Canada) at a rate of 68 mg NL- amended with 2 mLL- of Calcium-Magnesium supplement (0.0N0.0P0.0K3.0Ca1.6Mg; EZ-GRO Inc.) and 22.9 mg NL- of Organa ADD micronutrient supplement (2.0N0.0P0.0K; EZ-GRO Inc.), with a 20% leaching fraction. Other nutrient element concentrations in Organa ADD were (in mg L-): 100.0 Ca, 29851 Zn, 4892 Mn, 1239 B, 12.7 Mo, 2419 Cu and 2917 Fe.

Plant Culture and Treatments

Cannabis [Cannabis sativa L. WP:Med (Wappa)] cuttings were taken at a length of ≈13 cm and
with three fully-expanded leaves from stock plants. Cuttings were taken from the ends of axial limbs and cut at a 45˚ angle. Each cutting was rooted in a 5.7 cm wide, 5.7 cm tall peat-based pot (Jiffy Products N.B. Ltd., NB, Canada) containing Pro-Mix PG Organic growing substrate (Premier Tech ) and arranged in trays at a density of 266 plants/m2. The substrate was soaked in a solution of Spurt liquid organic fertilizer (2.0N0.0P 0.83K; EZ-GRO Inc.) at a rate that supplied 123 mg NL-.

The experiment was a full factorial completely randomized design with four factors (rooting hormone, leaf number, cutting position and leaf tip removal), two levels per factor and 10 replications per factor combination. For leaf number, cuttings had either one fully-expanded leaf removed (two leaves remaining) or were left with three leaves. For cutting position, cuttings
from terminal shoots were taken from either an apical position (node 10 and higher) or a basal position (below node 10). For the leaf tip removal treatment, a portion of leaf tips (≈1/3 of the leaf area) was removed from the fully expanded leaves or the leaves were left uncut. For the rooting hormone factor, the base (≈5 cm) of each stem was dipped in either 0.2% indole-3-butyric acid gel (synthetic rooting hormone; EZ-GRO Inc.) or in a 0.2% willow extract rooting gel (organic rooting hormone; EZ-GRO Inc.).

Propagation Environment

Trays were randomly arranged in a walk-in growth chamber (Conviron ATC60; Controlled Environments Ltd., Winnipeg, MB, Canada) and cuttings were misted with reverse osmosis (RO) water once, when they were placed in the chamber. From days 0 to 4 after cuttings were placed in the substrate (DAP), RH was maintained at 95% (s.d. 1.3), reduced to 80% (s.d 1.3%) for 5-8 DAP and to 60% (s.d. 1.5%) for 9-12 DAP. Temperature was maintained at 24 ˚C (s.d. 0.04˚C) (day/night) for the entire period. Fluorescent lighting (Philips Lighting, Markham, ON, Canada) was used to maintain an 18-hour photoperiod. Photosynthetically active radiation (PAR) at canopy level was maintained at 50 molm-s- (s.d 0.6 molm-s-) for 0-4 DAP 1-5, 80 molm-s- (s.d. 0.7 molm-s-) for 6-8 DAP and 115 molm-s- (s.d. 0.5 molm-s-) for 10-12 DAP.

Rooting Assessment and Harvest.

The bottom of the trays was observed daily from 7 DAP onwards for protruding roots, and cuttings were harvested at 12 DAP when approximately more than 50% of the cuttings showed visible roots at the bottom of the tray. Rooting success rate was measured on a binomial scale in which any visible adventitious root formation was considered rooted. Rooting success was calculated as the percentage of cuttings with roots in each treatment. Successfully rooted cuttings were assigned to either of two classifications based on degree of adventitious rooting: A root quality index (RQI) score of 1 or 2 was assigned by a third party without knowledge of the applied treatments based on a visual reference (Fig 1). Before RQI measurements, the substrate was washed from rooted cuttings with RO water and ratings were determined by a third party without knowledge of the applied treatments.

Statistical Analysis

Data were analyzed using JMP Statistical Discovery Version 13.0 (SAS Institute Inc., Cary, NC) at a Type 1 error rate of ≤ 0.05. Rooting success rate and RQI data were analysed assuming a binomial error distribution using a generalized linear model (GLM) and logit link function.
Stepwise regression with minimum AICC was used to remove non-significant interactive effects.
Chi-square contrasts were used to compare treatment means and interactive effects between treatments.

RESULTS

Cuttings under all treatment combinations had some degree of successful rooting. Rooting hormone had the greatest effect on both rooting success rate and root quality (Fig. 2, 3). The synthetic hormone delivered a 2.1 times higher rooting success rate (84% vs. 40%; χ= 39.0, P < 0.0001) and 1.6 times higher root quality (1.6 vs. 1.0; χ= 41.1, P < 0.0001) than the organic hormone. Removing leaf tips had the second greatest effect on rooting success rate. When leaf tips were removed, rooting success rate was lowered from 71% to 53% (χ = 9.8, P = 0.0018), though there was no effect on root quality. Leaf number had no effect on rooting success rate, but rooted cuttings with three leaves had 15% higher root quality than those with two (1.5 vs. 1.3; χ = 4.3, P = 0.038). Cutting position did not influence rooting success rate or root quality.
There was however, an interactive effect between cutting position and leaf tip removal on rooting success rate (Fig. 4). When leaf tips were removed, cuttings of basal origin had lower rooting success rate than apical cuttings (43% vs. 63%; χ = 5.7, P = 0.0169).

DISCUSSION

Rooting Hormone

The use of IBA led to markedly higher rooting success rate and root quality than the organic hormone. Similar success with IBA has been documented in other species propagated by stem cuttings (Al-Saqri and Alderson 1996; Saffari and Saffari 2012). In studies on the effects of centrifuged willow shoot extracts on willow and mung bean stem cuttings, willow extract application increased adventitious rooting in mung bean but had no effect in willow cuttings (Kawase 1964, 1970). The authors attributed the improved rooting in mung bean to a synergistic effect between indole-3-acetic acid (IAA) in the cuttings and two unknown root promoting fractions identified in willow extract. This synergistic effect was explored on adventitious rooting, also in bean cuttings, in Gesto et al. (1977) and attributed to the presence of the
compound catechol in willow extract. Kawase (1970) suggested that only cuttings with sufficient IAA, such as mung bean, would see improved rooting from the synergism. A recent evaluation on the effects of willow extract on olive stem cuttings (Al-Amad and Qrunfleh 2016) showed that, similar to willow cuttings, willow extract had no effect on adventitious rooting in olive
cuttings. The relatively poor rooting success rate of cannabis cuttings treated with willow extract in the present study could be attributed to a lack of IAA in the cuttings. Further study is required to measure IAA in cannabis cuttings and to further explore the synergistic effect between IAA and catechol on adventitious rooting. Also, more organic rooting hormones need to be explored and evaluated for cannabis propagation to provide alternatives for growers that choose organic production.

Leaf Number/ Leaf Tip Removal

Rooting success was similar between cuttings with two and three leaves, suggesting that two leaves may provide sufficient carbohydrates, auxin and rooting co-factors (Haissig 1974) for successful rooting in cannabis. In the propagation of stem cuttings, increased photosynthetic surface area and resultant carbohydrate supply generally increased rooting success rate until
another factor such as evapotranspiration stress became limiting (Davis and Potter 1989).
Cuttings with three leaves showed no signs of wilting or other indications of evapotranspiration stress. It is likely that both two- and three-leaf treatments exhibited little evapotranspiration-induced water stress in the stable, high humidity provided in this trial. It is estimated that under conditions of lower or less stable humidity, fewer leaves would deliver improved rooting success rate as these cutting would have a lower demand for humidity.

Notably, three leaves increased root quality over two. The observed quality improvement was likely caused by the additional carbohydrates, rooting co-factors, endogenous auxin (Haissig 1974) or a combination of these factors provided by the additional foliage. Further study is required to evaluate the effect of each of these factors and their interactions on cannabis stem cuttings to discern their relative importance. Based on this finding, it is recommended that cannabis cuttings be taken with three or potentially more leaves (to improve rooting quality) so long as humidity during propagation can be adequately maintained.

It was expected that cutting leaf tips would have a similar effect to reducing leaf number since the source and thus potentially the amount of photosynthetic material was manipulated; however, cutting leaf tips reduced rooting success and had no effect on rooting quality.

Both cutting leaf tips and leaf number alter surface area for evapotranspiration and photosynthesis; however, there was a notable difference in the effects of these treatments. Leaf cutting influenced rooting success rate while leaf number influenced root quality. Further study
is necessary to discern the reason for these differing effects. Based on these findings, it is recommended that leaf tips not be cut in cannabis cuttings; and, if less leaf material is desired to conserve space in the propagation environment or to prevent evapotranspiration stress, then fewer whole leaves be used instead.

Cutting Position

There was no indication that basal cuttings had improved rooting success rate or quality over apical cuttings. Similar results were found in stem cuttings of fever tea (Soundy et al. 2008) and may be attributed to the lack of distinct stages of maturation in these herbaceous plants in contrast to most hardwood species (Schreiber and Kawase 1975; Morgan and McWilliams 1976; Husen and Pal 2006). These findings suggested that in cannabis, cutting position does not play an important role in rooting.

Interactive effects. There was an interactive effect between leaf tip removal and cutting position.
Cuttings from basal positions with two leaves had lower rooting success rate than any other combination of these two factors. In general, basal cuttings had smaller leaves (general observation without measurement) than apical cuttings which, through cutting of leaf tips, were
left with less overall leaf surface area. The reduction in photosynthesis resulting from the smaller leaf area might have then resulted in the lower rooting success rate.

CONCLUSIONS

Type of rooting hormone strongly influenced the success and quality of adventitious rooting in cannabis cuttings, with the 0.2% IBA gel delivering higher rooting success rate than the 0.2% willow extract. Removing 30% of leaf tips from cuttings reduced rooting success rate and three leaves had higher root quality compared to two leaves without influencing rooting success rate.
Position of cutting on the stock plant did not influence either rooting success rate or root quality.
To achieve maximum rooting success and root quality, cuttings from either apical or basal positions should have at least three fully expanded, uncut leaves and be dipped in an IBA rooting hormone. If a reduction in leaf area is desired, either because high humidity cannot be
maintained or more airflow is desired in the propagation environment, then lowering leaf number to two fully expanded leaves is preferential to cutting leaf tips.









Fig 1. Visual scale for Rooting Quality Index (RQI) assessment of cannabis cuttings.

Fig 2. Rooting success rate of cannabis cuttings (means SEM; n = 80). Cut leaves had about 30% of leaf tips were removed. Cuttings were from terminal shoots with apical cuttings taken from ≥ node 10 and basal cutting from < node 10. The synthetic rooting hormone was a 0.2% indole-3-butyric (IBA) acid gel and the organic was a 0.2% willow extract gel. Bars within each factor (e.g. leaf number) bearing different letters are significantly different at P < 0.05 using chi squared contrasts.

Fig 3. Root Quality Index of cannabis cuttings (means SEM; n = 80). Cut leaves had about 30% of leaf tips were removed. Cuttings were from terminal shoots with apical cuttings taken from ≥ node 10 and basal cutting from < node 10. The synthetic rooting hormone was a 0.2%
indole-3-butyric (IBA) acid gel and the organic was a 0.2% willow extract gel. Bars within each factor (e.g. leaf number) bearing different letters are significantly different at P < 0.05 using chi squared contrasts.

Fig 4. Rooting success rate of cannabis cuttings (means SEM; n = 40). Cut leaves had about 30% of leaf tips were removed. Cuttings were from terminal shoots with apical cuttings taken from ≥ node 10 and basal cutting from < node 10. Treatments with different letters are
significantly different at P < 0.05 using Chi Squared contrasts.
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Last edited by Efesto; 01-04-2019 at 01:43 PM.
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Old 01-04-2019, 03:02 AM
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I cut my clones, hand strip what I want off then use cloning gel and put them in cups of cold water. We have about a 96% success rate this way for the last 14 1/2 years and counting. But I am fairly lazy
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Old 01-04-2019, 01:48 PM
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Quote:
Originally Posted by Dandaweedman View Post
I cut my clones, hand strip what I want off then use cloning gel and put them in cups of cold water. We have about a 96% success rate this way for the last 14 1/2 years and counting. But I am fairly lazy
96%... there is still room for improvement...lol
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Old 01-04-2019, 03:34 PM
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Always room for improvement for sure.
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Old 01-04-2019, 05:04 PM
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I don't use any hormones most of the time, occasionally willow water when I think to harvest willow fronds. I don't snip the leaf tips but I pull them off. I do that because I was a salad prep guy and the difference between lettuce pulled apart and cut apart is quite large, with the pulled lasting days longer.

I'm pretty sure my rate is somewhere around 95%, but I don't keep records. I'm not worried about my rates.
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Old 01-04-2019, 11:17 PM
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As long as you don't lose them that's all that matters. I'm used to keeping data on me babies. excellent in future endeavours.
I used to make my own willow extracts for cloning. Lots of willows here but I got lazy
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